This is a competing continuation application of our SBIR Phase II project entitled "Effective and Non-Toxic Antagonist to Heparin and LMWH". Extremely promising results and outstanding productivity have been attained from the previous Phase II research, as reflected by the publication of 12 peer-reviewed manuscripts and 2 US patents. Both in vitro and animal testing demonstrated that the low molecular weight protamine (LMWP) under development not only completely neutralized the activities of both heparin and LMWH, but also did not elicit any adverse responses. In addition, the chain-shortened LMWP was shown to be deprived of both antigenicity and immunogenicity;two primary toxic effects of protamine. Furthermore, preliminary studies in diabetic rats indicated that LMWP could be formulated with insulin to yield a long-lasting blood glucose control without displaying the commonly observed protamine reactions that masquerade as insulin allergy. The main objective of this competing continuation application is to carry out the pre-clinical safety studies, required by FDA for Investigational New Drug (IND), to develop LMWP as a non-toxic clinical substitute for protamine in all of its possible applications (e.g. heparin reversal, insulin formulations), for all patients, without any concern of protamine-associated toxic effects. The six highly integrated specific aims are to: [1] design a packed-bed reactor (PBR) flow system that allows for GMP-guided manufacturing of LMWP with the highest purity and yield;[2] assess the toxic and pharmacologic effects of LMWP via in vitro and in vivo animal testing;[3] determine the acute toxicity of the LMWP product in both rats and rabbits;[4] establish the safety pharmacology profile of LMWP (Note: Aim [2] through Aim [4] will be conducted by following the FDA Guidelines for IND);[5] evaluate the efficacy and toxicity of LMWP in protamine/LMWP-sensitized rats;and [6] evaluate the efficacy and toxicity LMWP/insulin formulations in a diabetic rats. Success of these aims will lead to LMWP becoming a whole-sale clinical protamine substitute. The annual consumption of protamine involved in heparin reversal is estimated to be over 10 metric tons. The quantity of protamine used in insulin formulations is believed to be even several folds higher. Therefore, the predicted market for LMWP is very significant, and the impact and true value of the proposed LMWP technology is far-reaching.